WO2019089584A2 - Amphiphiles peptidiques triblocs, micelles et leurs méthodes d'utilisation - Google Patents

Amphiphiles peptidiques triblocs, micelles et leurs méthodes d'utilisation Download PDF

Info

Publication number
WO2019089584A2
WO2019089584A2 PCT/US2018/058200 US2018058200W WO2019089584A2 WO 2019089584 A2 WO2019089584 A2 WO 2019089584A2 US 2018058200 W US2018058200 W US 2018058200W WO 2019089584 A2 WO2019089584 A2 WO 2019089584A2
Authority
WO
WIPO (PCT)
Prior art keywords
peptide
acid
triblock
alternative
lipid
Prior art date
Application number
PCT/US2018/058200
Other languages
English (en)
Other versions
WO2019089584A3 (fr
Inventor
Bret ULERY
Rui Zhang
Caitlin LEEPER
Josiah SMITH
Logan MORTON
Original Assignee
The Curators Of The University Of Missouri
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Curators Of The University Of Missouri filed Critical The Curators Of The University Of Missouri
Priority to US16/760,329 priority Critical patent/US20200339661A1/en
Publication of WO2019089584A2 publication Critical patent/WO2019089584A2/fr
Publication of WO2019089584A3 publication Critical patent/WO2019089584A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/76—Albumins
    • C07K14/77—Ovalbumin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/57563—Vasoactive intestinal peptide [VIP]; Related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • compositions such as vaccines and immunomodulatory therapies, comprising a triblock peptide and methods of use.
  • PAMs peptide amphiphile micelles
  • the triblock peptide is of the formula:
  • A is a lipid moiety
  • B and C are independently a peptide block or a zwitterion-like block.
  • compositions comprising a triblock peptide of the formula:
  • A is a lipid moiety; and B and C are independently a peptide block or a zwitterion-like block.
  • the triblock peptides may be arranged in micelles in a pharmaceutically acceptable carrier.
  • the pharmaceutical composition may be a vaccine composition, which optionally may additionally comprise an adjuvant.
  • An additional aspect of the present disclosure is directed to a method of treating a disease or condition in a subject.
  • the method comprises administering a therapeutically effective amount of a triblock peptide to the subject, wherein the triblock peptide is of the formula:
  • A is a lipid moiety; and B and C are independently a peptide block or a zwitterion-like block.
  • the triblock peptides may be arranged in micelles in a pharmaceutically acceptable carrier.
  • the pharmaceutical composition may be a vaccine composition, which optionally may additionally comprise an adjuvant.
  • FIG. 1 depicts exemplary ABC triblock peptide amphiphiles of the present invention and exemplary lipids and zwitterion-like blocks.
  • FIG. 2 depicts an exemplary micelle vaccine of the present invention carrying a lipid based adjuvant.
  • FIG. 3 depicts an exemplary micelle vaccine of the present invention carrying a nucleic acid based adjuvant.
  • FIG. 4 depicts a micrograph of PalmK-OVABT peptide amphiphile showing it self-assembles into cylindrical micelles in water.
  • FIG. 8 depicts the disassociation of higher order micellar structures dependent on charge status changes in the zwitterion-like block due to lysine protonation or glutamic acid deprotonation.
  • the formation or disassociation of aggregated twine-like micellar structures is dependent on the zwitterion-like block charge status.
  • charge complexation across multiple micelles due to local dipole moments is possible.
  • the zwitterion-like block possesses a significant amount of non-charged glutamic acids or lysines, respectively, yielding electrostatically repulsive peptide segments that prevent complex micelle aggregation.
  • FIG. 9A, FIG. 9B, and FIG. 9C depict micrographs of PalmK-0 VABT at three different pHs (2 (FIG. 9A), 7 (FIG. 9B), and 11 (FIG. 9C)) showing no obvious morphological changes as a function of altering pH indicating the importance of the zwitterion-like region in complex micelle formation.
  • FIG. 10A, FIG. 10B, FIG. IOC, FIG. 10D, and FIG. 10E depict PalmK-(EK) 4 - OVABT complex micelle aggregation as pH-dependent. PalmK-(EK) 4 -OVABT was solubilized in different pH solutions to investigate the impact of this parameter on micelle formation as assessed by negative stain TEM ((FIG. 10A) 3, (FIG. 10B) 7, and (FIG. IOC) 11). The influence pH has on peptide secondary structure was also evaluated by (FIG. 10D) obtaining the CD spectra and using it to (FIG. 10E) estimate secondary structure.
  • FIG. 11 A, FIG. 11B, and FIG. 11C depict micrographs at three different magnifications (FIG. 11A - 3,000 x, FIG. 11B - 8,000 x, FIG. 11C - 20,000 x) showing PalmK-(EK)4-OVABT forms both thread-like and braided micelles at neutral pH.
  • FIG. 12A and FIG. 12B depict a PalmK-OVA B T-(KE) 4 micrograph (FIG. 12A) and a PalmK-(EK) 4 -OVABT micrograph (FIG. 12B) with micellar diameter measurements of 41.38 nm, 29.71 nm and 115.67 nm.
  • FIG. 13 depicts micelle braiding dependent on the zwitteri on-like block location in PalmK-OVABT-(KE) 4 and PalmK-(EK) 4 -OVABT.
  • Micelle aggregation and its effect on the resulting final nanomaterial structure is dependent on how easily dipole surface charges can associate across micelles.
  • unmatched dipole surface charges on the twine-like micelles further drive the formation of braided micelles via electrostatic complexation.
  • PalmK-(EK) 4 -OVABT is expected to form flowerlike micelles to best position the zwitteri on-like block in the corona, which significantly reduces amphiphile fluidity within the individual micelle. This limits the capacity for twining to match most or all charge dipoles allowing for the remaining unmated regions to further complex twines into braids.
  • FIG. 14A, FIG. 14B, FIG. 14C, and FIG. 14D depict electrostatically driven micelle aggregation as insensitive to physiological ion content.
  • Negative-stain TEM revealed that PalmK-OVABT-(KE) 4 and PalmK-(EK) 4 -OVABT form twinelike and braidlike micelles, respectively, regardless of whether the amphiphiles were exposed to low salt concentration (i.e., ddH 2 0) or high salt concentration (i.e., PBS). Palm 2 K-OVA B T-(KE) 4 in ddH 2 0 (FIG. 14A) and in PBS (FIG. 14B) and PalmK-(EK) 4 -OVA B T in ddH 2 0 (FIG. 14C) and in PBS (FIG. 14D)
  • low salt concentration i.e., ddH 2 0
  • PBS high salt concentration
  • FIG. 15A, FIG. 15B, FIG. 15C, and FIG. 15D depict CD spectra of four different PA formulations (FIG. 15A - PalmK-(EK) 4 -OVA B T, FIG. 15B - Palm 2 K-(EK) 4 - OVABT, FIG. 15C - PalmK-OVA B T-(KE) 4 , and FIG. 15D - Palm 2 K-OVA B T-(KE) 4 ) in either ddH 2 0 (pH adjusted to 7) or in PBS. Certain increases in ion concentration do not significantly alter micellar secondary structure.
  • the CD spectra under 200 nm is variable in PBS solution due the high salt concentration interrupting absorbance at lower wavelengths.
  • FIG. 16A, FIG. 16B, FIG. 16C, and FIG. 16D depict micelle shape and aggregation as dependent on hydrophobic content and zwitteri on-like block location.
  • OVABT amphiphile chemistries ((FIG. 16A) PalmK-OVA B T-(KE) 4 , (FIG. 16B) Palm 2 K-OVA B T-(KE) 4 , (FIG. 16C) PalmK-(EK) 4 -OVA B T, and (FIG. 16D) Palm 2 K-(EK) 4 - OVABT) yielded significantly different micellar structures (i.e., (FIG. 16A) twines, (FIG.
  • FIG. 17 depicts a micrograph of Palm 2 K-OVABT peptide amphiphile showing it self-assembles into spheres and cylindrical micelles in water.
  • FIG. 18A, FIG. 18B, FIG. 18C, FIG. 18D, FIG. 18E, and FIG. 18F depict micrographs of Palm 2 K-OVA B T-(KE)4 (FIG. 18A, FIG. 18B, and FIG. 18C) and Palm 2 K- (EK) 4 -OVABT (FIG. 18D, FIG. 18E, and FIG. 18F) in ddH20 at three different pHs. While the micelles dissociated from one another at acidic or basic conditions, no other morphological changes were observed. This result is believed to be caused by hydrophobic interactions being more dominant than the electrostatic interactions.
  • FIG. 19A, FIG. 19B, FIG. 19C, and FIG. 19D depict CD spectra of Palm 2 K- OVABT-(KE) 4 (FIG. 19A) and Palm 2 K-(EK) 4 -OVA B T (FIG. 19C) with their respective secondary structure estimation (FIG. 19B and FIG. 19D, respectively).
  • FIG. 20A and FIG. 20B depict micrographs of Palm 2 K-OVA B T-(KE) 4 (FIG.
  • Palm 2 K-(EK) 4 -OVA B T (FIG. 20B) in ddH 2 0 (pH adjusted to 7).
  • Significant aggregation of Palm 2 K-(EK) 4 -OVABT was observed whereas only slight aggregation was shown for Palm 2 K-OVABT-(KE) 4 .
  • double tail PAs have similar fluidity differences as their single tail counterparts so Palm2K-(EK)4-OVABT is less able to match dipoles within individual micelles allowing for greater multiple micelle complexation.
  • FIG. 21A, FIG. 21B, and FIG. 21C depict micrographs of Palm 2 K-OVA BT (FIG. 21A, FIG. 21B, and FIG. 21C) at three different pHs (2, 7, and 11), showing no obvious morphological changes as a function of altering Ph, indicating the importance of the zwitteri on- like region in complex micelle formation.
  • FIG. 22A, FIG. 22B, FIG. 22C, and FIG. 22D depict micelle shape and aggregation as application-specific peptide insensitive.
  • OVAcytoT amphiphile chemistries ((FIG. 22A) PalmK-OVAcytoT- KE (FIG. 22B) Palm 2 K-OVAc y toT-(KE) 4 , (FIG. 22C) PalmK- (EK) 4 -OVAcytoT, and (FIG. 22D) Palm 2 K-(EK)4-OVAcytoT) similar to the OVABT peptide amphiphiles yielded essentially identical structures (i.e., (FIG. 22 ⁇ ) twines, (FIG.
  • FIG. 22B depicts PalmK-OVA B T-(KE) 4 CMC determination at different pH values.
  • FIG. 24 depicts Palm 2 K-OVA B T-(KE)4 CMC determination at different pH values.
  • FIG. 25 depicts PalmK-(KE) 4 -OVA B T CMC determination at different pH values.
  • FIG. 26 depicts Palm 2 K-(KE)4-OVA B T CMC determination at different pH values.
  • FIG. 27 depicts PalmK-OVAcytoT-(KE) 4 CMC determination at different pH values.
  • FIG. 28 depicts Palm 2 K-OVA cy toT-(KE) 4 CMC determination at different pH values.
  • FIG. 29 depicts PalmK-(KE) 4 -OVA cyt oT CMC determination at different pH values.
  • FIG. 30 depicts Palm 2 K-(KE) 4 -OVA cyt0 T CMC determination at different pH values.
  • FIG. 31A and FIG. 31B depict HPLC photo diode array detection (FIG. 31A) and mass spectrometry analysis (FIG. 31B) of PalmK-OVA B T-(KE) 4 .
  • FIG. 32A and FIG. 32B depicts HPLC photo diode array detection (FIG. 32A) and mass spectrometry analysis (FIG. 32B) of Palm 2 K-OVA B T-(KE) 4 .
  • FIG. 33A and FIG. 33B depict HPLC photo diode array detection (FIG. 33A) and mass spectrometry analysis (FIG. 33B) of PalmK-(EK) 4 -OVA B T.
  • FIG. 34A and FIG. 34B depict HPLC photo diode array detection (FIG. 34A) and mass spectrometry analysis (FIG. 34B) of Palm 2 K-OVA B T-(KE) 4 .
  • FIG. 35A and FIG. 35B depict HPLC photo diode array detection (FIG. 35A) and mass spectrometry analysis (FIG. 35B) of PalmK-OVA cy toT-(KE) 4 .
  • FIG. 36A and FIG. 36B depict HPLC photo diode array detection (FIG. 36A) and mass spectrometry analysis (FIG. 36B) of Palm 2 K-OVA cy toT-(KE) 4 .
  • FIG. 37A and FIG. 37B depict HPLC photo diode array detection (FIG. 37A) and mass spectrometry analysis (FIG. 37B) of PalmK-(EK) 4 -OVA cy toT.
  • FIG. 38A and FIG. 38B depict HPLC photo diode array detection (FIG. 38A) and mass spectrometry analysis (FIG. 38B) of Palm 2 K-(EK) 4 -OVA cy toT.
  • FIG. 39A pVIPA and FIG. 39B) pzVIPA formed cylindrical micelles and braided micelles, respectively.
  • FIG. 40 A and FIG. 40B depict peptide secondary structure analysis.
  • the influence lipidation and zwitterion-like peptide block inclusion had on peptide secondary structure was evaluated by obtaining the CD spectra (FIG. 40A) and using it to estimate secondary structure (FIG. 40B).
  • FIG. 41A, FIG. 41B, and FIG. 41C depict anti-inflammatory effects of different VIP formulations.
  • T F-a secretion from M0s (FIG. 41A) or DCs (FIG. 41B) as well as CD86 expression from DCs (FIG. 41C) were evaluated.
  • LPS greatly increased each of these inflammatory correlates which were diminished to variable extents due to the presence of different concentrations and presentations of VIP.
  • groups that possess different letters have statistically significant differences in mean (p ⁇ 0.05) whereas those that possess the same letter are similar (p > 0.05).
  • FIG. 42A, FIG. 42B, FIG. 42C, FIG. 42D, FIG. 42E, and FIG. 42F depict inflammatory signal regulation of the lipid moiety (i.e. palmitic acid - Palm).
  • FIG. 43A, FIG. 43B, and FIG. 43C depict immunoregulatory effects of different VIP formulations.
  • the secretion of CCL22 from immature DCs (FIG. 43A) and mature DCs (FIG. 43B) as well as the CD86 expression on DCs (FIG. 43C) was evaluated.
  • the production of T reg recruiting CCL22 was enhanced for some VIP AM formulations.
  • pVIPA significantly increased CD86 expression on immature DCs while the other two VIP formulations did not enhance CD86 expression.
  • groups that possess different letters have statistically significant differences in mean (p ⁇ 0.05) whereas those that possess the same letter are similar (p > 0.05).
  • FIG. 44A and FIG. 44B depict CCL22 induction effects of the lipid moiety (i.e. palmitic acid - Palm).
  • the secretion of CCL22 from immature DCs (FIG. 44A) and mature DCs (FIG. 44B) was evaluated. Palm was found to have no impact on CCL22 induction regardless of DC maturation state.
  • groups that possess different letters have statistically significant differences in mean (p ⁇ 0.05) whereas those that possess the same letter are similar (p > 0.05).
  • One aspect of the present invention is directed to triblock peptides comprising a lipid moiety, a peptide block and a zwitterion-like block.
  • the peptides of the invention are useful in forming micelles. It has been found that adding a zwitterion-like block to a peptide- lipid amphiphile provides benefits over use of the peptide-lipid amphiphiles alone.
  • compositions comprising the triblock peptides of the present in invention arranged in micelles in a pharmaceutically acceptable carrier.
  • the physical properties, such as size, shape and charge of the resulting micelles may be modified by the selection and order of the lipid, peptide and zwitterion-like components of the triblock peptide. These properties are closely related to micelle immunogenicity.
  • the pharmaceutical compositions of the present invention are vaccine compositions.
  • the peptide block of the vaccine may comprise the immunogenic peptide epitope of the target pathogen.
  • the vaccine compositions may further comprise an adjuvant, which may be carried by the vaccine micelles.
  • Another aspect of the invention is directed to methods of using the triblock peptides, pharmaceutical compositions and vaccine compositions of the present invention to treat a disease or condition in a subject.
  • the present disclosure is directed to a triblock peptide of the formula:
  • A is a lipid moiety
  • B and C are independently a peptide block or a zwitterion-like block.
  • the unique ABC triblock peptides of the present invention are capable of forming complex nanostructures. As noted above, this is an improvement over traditional peptide amphiphiles that do not include a zwitterion-like block. These complex nanostructures are discussed in more detail in section 11(A), below. Exemplary triblock peptides of the present invention are depicted in FIG. 1, along with exemplary lipids and zwitterion-like blocks.
  • the lipid moiety may be a C 2 - C 3 8 saturated fatty acid, for example, C 2 , C 4 , C 6 , C 8 , C10, Ci 2 , C14, Ci6, Ci 8 , C 2 o, C 22 , C 24 , C 26 , C 28 , C30, C 32 , C 34 , C 36 , and C 38 any number or range of carbon atoms there between.
  • the lipid moiety may include linkers (e.g., lysine, glutamic acid, aspartic acid, citric acid, and glycerol) that allow for the attachment of one or two fatty acids, or even more, such as three, four, or any number up to eight fatty acids.
  • Suitable lipid moieties include, without limit, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecanoic acid, palmitic acid, margaric acid, stearic acid, nonadecylic acid, arachidic acid, heneicosylic acid, behenic acid, tricosylic acid, lignoceric acid, pentacosylic acid, cerotic acid, heptacosylic acid, montanic acid, nonacosylic acid, melissic acid, henatriacontylic acid, lacceroic acid, psyllic acid, geddic acid, ceroplastic acid, hexatriacontylic acid, heptatriacontanoic acid, octatricontanoic acid,
  • the lipid moiety may also be a C 2 - C 38 unsaturated fatty acid, as di cussed above with respect to saturated fatty acids, containing at least one carbon-carbon cis or trans double bond, for example Ci 8:3 , Ci 8:4 , C 2 o:5, C 22:6 , Ci 8:2 , C 2 o: 3 , C 2 o: 4 , C 22:4 , Ci6:i, Ci 8: i, C 2 o:i, C 22: i, and C 24: i. Similar to saturated fatty acids, the lipid moiety can consist of one to eight unsaturated fatty acids held together by suitable linker molecules.
  • Suitable lipid moieties include, without limit, a-linolenic acid, stearidonic acid, eicosapentaenoic acid, docosahexaenoic acid, linoleic acid, linolelaidic acid, ⁇ -linolenic acid, dihomo-y-linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid, erucic acid, nervonic acid, mead acid, and their combinations.
  • the lipid moiety can also be a bioactive lipid containing anywhere from one to eight saturated and/or unsaturated fatty acids (C 2 - C 38 , as dicussed above with respect to saturated fatty acids), linked together.
  • Suitable bioactive lipid moieties include, without limit, valproic acid, monophosphoryl lipid A (MPLA) and its analogs, dipalmityolcysteinylserinyltetralysine (P 2 CSK 4 ), squalamine and its analogs, squalene and its analogs, leukotriene B 4 , prostaglandin E 2 , thromboxane A 2 , prostacyclin I 2 , phosphatidylserine, phosphatidylinositol, lysophosphatidic acid, sphingosine- 1-phosphate, N-arrachidonylethanolamine, 2-arachidonylglycerol, N-palmitoylethanolamine, eicos
  • the peptide block may be a peptide comprised of anywhere from one to fifty amino acids in length.
  • the lipid moiety may include linkers (e.g., lysine, glutamic acid, aspartic acid, citric acid, and glycerol) that allow for the attachment of one or two peptides, or even more, such as three, four, or any number up to eight peptides.
  • the resulting peptide block may range from 75 g/mol to 80,000 g/mol in molecular weight.
  • the term peptide refers to one or more linked amino acids, which may comprise, without limit, a portion of a protein, a peptide epitope or a complete protein.
  • the peptide block comprises a peptide to treat or target a disease or condition.
  • the peptide block comprises an immunogenic peptide epitope which can be used to produce micelle vaccines.
  • Another embodiment includes a peptide comprised of an immunoactive peptide which can be used to make immunomodulatory micelles.
  • vaccines and immunotherapeutics would be safe, cost and time effective, stable at room temperature and have low immugenicity.
  • Other suitable peptides may include peptides to target and/or attack cancer cells (anti-cancer peptides).
  • Suitable peptide blocks include, without limit, ovalbumin and vasoactive intestinal peptide, for example OVABT (ESLKISQAVHAAHAEINEAGRE) (SEQ ID NO: 1), OVAcytoT (EQLESIINFEKLTE) (SEQ ID NO: 2), as well as the immunogenic peptides, immunoactive peptides and anti -cancer peptides set forth in Tables 1, 2 and 3, below.
  • OVABT ESLKISQAVHAAHAEINEAGRE
  • OVAcytoT EQLESIINFEKLTE
  • immunogenic peptides, immunoactive peptides and anti -cancer peptides set forth in Tables 1, 2 and 3, below.
  • N/A KLDVGNAEV 10.1 158/1078-0432.CCR-10- 2614 N/A YLMDTSGKV 10.1158/1078-0432.CCR-10- 2614
  • the peptides are selected for treatment of influenza.
  • Exemplary peptides include, without limit, the peptides listed below:
  • Macrophage targeting micelles that also enhance micelle-associated anti-inflammatory bioactivity
  • the peptide blocks may be synthesized using techniques known to those of skill in the art. Such methods include peptide coupling reagents, such as carbodiimides, aminium/uranium and phosphonium salts, solid supports, such as gel-type supports, surface-type supports, and composites, protecting group schemes, such as Boc/Bzl, Fmoc/tBu, benzyloxy-carbonyl, alloc, and regioselective disulfide bond formation, microwave-assisted synthesis, and on- and off-resin cyclization. Such methods may be used in combination with others, such as solid phase synthesis using Fmoc chemistry.
  • peptide coupling reagents such as carbodiimides, aminium/uranium and phosphonium salts
  • solid supports such as gel-type supports, surface-type supports, and composites
  • protecting group schemes such as Boc/Bzl, Fmoc/tBu, benzyloxy-carbonyl, all
  • the zwitteri on-like block is generally comprised of a peptide comprised of a combination of positively charged, negatively charged, and neutral amino acids two to fifty amino acids in length that yields some local regions of positive and negative charge that can facilitate complexation.
  • Suitable zwitterion-like blocks may include, without limit, ( ⁇ ) ⁇ where ⁇ , ⁇ , and ⁇ consists of a positively charged amino acid (K - lysine and/or R - arginine), a negatively charge amino acid (E - glutamic acid and/or D - aspartic acid), and a neutral amino acid (G - glycine and/or A - alanine) and X, Y, and Z, can be any number from 0 - 50, and B can be any number from 0 - 1.
  • the zwitterion-like block may include linkers (e.g., lysine, glutamic acid, aspartic acid, citric acid, and glycerol) that allow for the attachment of one or two zwitterion-like peptides, or even more such as three, four, or any number up to eight zwitterion-like peptides.
  • linkers e.g., lysine, glutamic acid, aspartic acid, citric acid, and glycerol
  • the resulting zwitterion-like block may range from 200 g/mol to 60,000 g/mol in molecular weight.
  • Suitable zwitterion-like blocks may include, without limit, (KXEYGZ)B, (KXGYEZ)B, (E X K Y GZ)B, (E X GYK z )B, (GXK Y E z )B, (GXE Y K z )B, (RXE Y GZ)B, (RXG Y E z )B, (E X R Y GZ)B, (E X G Y RZ)B, (GXR Y E z )B, (GXE Y R z )B, (K X D Y GZ)B, (K X G Y D z )B, (DXK Y GZ)B, (DXG Y K z )B, (G X K Y D z )B, (RXD Y G z )B, (RXG Y G z )B, (RXG
  • the zwitterion-like blocks may be (KE)x,(EG)x, (KA)x, (KG)x, including wherein X is 4.
  • blocks A, B, and C may be arranged lipid-peptide- zwitterion, lipid-zwitterion-peptide, peptide-lipid-zwitterion, peptide-zwitterion-lipid, zwitterion-lipid-peptide, or zwitterion-peptide-lipid.
  • Each of blocks A, B and C may contain complex, multiple component moieties as discussed above.
  • the peptide block may be synthesized using techniques known to those of skill in the art.
  • the peptide block may be synthesized using solid phase synthesis using Fmoc chemistry.
  • the Fmoc protecting group may be removed using piperidine in dimethylformamide (DMF).
  • the peptide block may be modified by orthogonal deprotection with the aid of either Fmoc-Lys(Fmoc)-OH or Fmoc-Lys(Dde)-OH conjugated to the N terminus of the peptides depending on whether single or multiple lipid moiety conjugation is desired.
  • the two lysine chemistries can be used singularly or in multiple combinations to create from one to eight chemical handles on which lipids can be conjugated.
  • a peptide can be initiated with either Fmoc-Lys(Fmoc)- OH or Fmoc-Lys(Dde)-OH on the C terminus for which a single or multiple amino acids can be included to allow from one to eight chemical handles on which peptides can be built.
  • Fmoc-Lys(Fmoc)- OH or Fmoc-Lys(Dde)-OH on the C terminus for which a single or multiple amino acids can be included to allow from one to eight chemical handles on which peptides can be built.
  • the same approach can be taken for the zwitterion-like block.
  • composition comprising a triblock peptide of the formula:
  • A is a lipid moiety
  • B and C are independently a peptide block or a zwitterion-like block
  • a pharmaceutically acceptable carrier including any of the peptides discussed above.
  • the triblock peptides are arranged in a micelle.
  • the triblock peptide may self-assemble into a micelle in the pharmaceutically acceptable carrier or other liquid.
  • the unique ABC triblock peptides of the present invention are capable of forming complex nanostructures. These include second-order (i.e., twines) and third-order (i.e., braids) micellar aggregates which are driven by intermolecular electrostatic complexation facilitated by the presence of a zwitterion-like peptide block. These interactions were found to be complementary of hydrophobically driven micellar self-assembly conveyed by the use of a fatty acid-based lipid provided these intramolecular forces were similar in strength.
  • the present invention leverages zwitterion-like peptides and their electrostatic interactions to achieve unique, environmentally sensitive micelle aggregates comprised of a variety of interesting and complex architectures.
  • zwitterion-like region has several advantages over comparable systems including their biocompatibility, solubility, and synthetic flexibility.
  • Previous research has shown that zwitterionic materials are quite hydrophilic which can contribute to enhance micelle stability as well as favorable interactions with biological systems. Because of block length and PA location choice, considerable control over micelle size, aggregate shape, peptide secondary structure, and stimuli sensitivity can be achieved. When coupled with the fact that these facets were found to be application-specific peptide independent, triblock PAs have the potential to function as a unique platform technology for use in a wide variety of biomedical subfields.
  • Electrostatic interactions can act as a complementary driving force to hydrophobic self-assembly facilitating the formation of aggregated micellar structures.
  • triblock peptide amphiphiles with carefully selected components are believed to be able to yield a wide array of self-assembled nanostructures in solution.
  • Multiple approaches such as changing block sequence, block ratio, and solvent conditions can possibly further alter their structure similarly to other comparable systems.
  • the individual micelles are spherical, cylindrical, or worm-like. These structures can range from 4 nm in diameter for small spherical micelles to 100 ⁇ in length for worm -like micelles.
  • micelles bearing a zwitterion-like block undergo electrostatic complexation yielding higher-order structures bearing complex architectures including, without limit, clusters, twines, braids, and nets. These structures can range from 10 nm in diameter for small cluster aggregates to 100 ⁇ in each dimension for net-like aggregates.
  • Micelle morphology may be determined using techniques known to those of skill in the art. Such techniques include transmission electron microscopy (TEM). Micelle secondary structure may be determined using techniques known to those of skill in the art. Such techniques include circular dichroism (CD).
  • TEM transmission electron microscopy
  • CD circular dichroism
  • the peptides confined within the micelles may form secondary structures including, without limit, a-helix, ⁇ -sheet, triple helix, 3-10 helix, and random coil.
  • pH influences such secondary structure formation. At neutral and basic pH conditions, a ⁇ -sheet conformation has been observed and at acidic pH conditions, a random coil and some a-helix conformations has been observed for certain formulations.
  • the inclusion of a bioactive peptide and zwitterion-like block has direct impact on micelle charge. Based on zeta potential measurements, the charge of triblock peptide amphiphile micelles can be from -60 mV to 60 mV.
  • the critical micelle concentration (CMC) may be determined using techniques known to those of skill in the art. Such methods include l,6-diphenyl-l,3,5-hexatriene (DPH) fluorescence. In some embodiments, the critical micelle concentration (CMC) may be from about 0.05 ⁇ to about 50 ⁇ .
  • the critical micelle concentration (CMC) may be about 0.05 ⁇ , about 0.1 ⁇ , about 0.15 ⁇ , about 0.2 ⁇ , about 0.25 ⁇ , about 0.3 ⁇ , about 0.35 ⁇ , about 0.4 ⁇ , about 0.45 ⁇ , about 0.5 ⁇ , about 0.55 ⁇ , about 0.6 ⁇ , about 0.65 ⁇ , about 0.7 ⁇ , about 0.75 ⁇ , about 0.8 ⁇ , about 0.85 ⁇ , about 0.9 ⁇ , about 1.0 ⁇ , about 1.25 ⁇ , about 1.5 ⁇ , about 1.75 ⁇ , about 2.0 ⁇ , about 2.25 ⁇ , about 2.5 ⁇ , about 2.75 ⁇ , about 3.0 ⁇ , about 3.25 ⁇ , about 3.5 ⁇ , about 3.75 ⁇ , about 4.0 ⁇ , about 4.25 ⁇ , about 4.5 ⁇ , about 4.75 ⁇ , about 5.0 ⁇ , about 5.25 ⁇ , about 5.5 ⁇ , about 5.75 ⁇ , about 6.0 ⁇ , about 6.25 ⁇ , about
  • the pharmaceutical composition may be a vaccine composition.
  • the micelles formed from tribock peptides of the invention can be tailored for the vaccine composition. Vaccine size and shape can determine its ability to travel to lymph node and cell uptake ability. Vaccine charge may affect its ability to interact with cells.
  • the morphology and charge of the micelles comprising the triblock peptide of the invention can be modulated to produce morphologies targeted for the intended use. Micelle morphology is discussed in more detail in Section 11(A) above and the Examples. Exemplary immunogenic peptides are discussed in Section I, above.
  • the vaccine composition may further include a pharmaceutically acceptable excipient such as a suitable adjuvant.
  • a pharmaceutically acceptable excipient such as a suitable adjuvant.
  • Adjuvants can create an antigen depot or display danger signals to the host to generate stronger protection.
  • the micelle vaccines of the present invention can be used in combination with adjuvants to enhance the benefits of the vaccine.
  • the adjuvant may include, without limit, an analgesic adjuvant, an inorganic compound, a mineral oil, a bacterial product, a delivery system, a cytokine, a food-based oil, a nonbacterial organic compound, an oligonucleotide, or a plant based saponin.
  • suitable adjuvants may include, without limit, an aluminium salt such as aluminium hydroxide or aluminium phosphate, but may also be a salt of calcium, iron or zinc, or may be an insoluble suspension of acylated tyrosine, or acylated sugars, or may be cationically or anionically derivatised saccharides, polyphosphazenes, biodegradable microspheres, ceramide, monophosphoryl lipid A (MPLA), lipid A derivatives (e.g., of reduced toxicity), 3-O-deacylated MPL [3D-MPL], quit A, Saponin, QS21, Freund's Incomplete Adjuvant (Difco Laboratories, Detroit, Mich.), Merck Adjuvant 65 (Merck and Company, Inc., Rahway, N.J.), AS-2 (Smith-Kline Beecham, Philadelphia, Pa.), CpG oligonucleotides, poly(LC), bioadhesives and mucoadhesives
  • Human immunomodulators suitable for use as adjuvants in the invention include cytokines such as interleukins (e.g., IL-1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12, etc.), macrophage colony stimulating factor (M-CSF), tumour necrosis factor (TNF), granulocyte, macrophage colony stimulating factor (GM-CSF) may also be used as adjuvants.
  • the micelle vaccines can be used as an adjuvant delivery vehicle.
  • a lipid based adjuvant such as a ceramide or MPLA, or a nucleic acid based adjuvant, such as CpG-ODN or Poly(LC) can be carried by the micelle vaccine, as depicted in FIG. 2 and FIG. 3.
  • a lipid based adjuvant such as ceramide or MPLA
  • a nucleic acid based adjuvant such as CpG-ODN or Poly(LC)
  • the adjuvants may be carried by electrostatic interactions.
  • compositions of the present invention will typically, in addition to the antigenic and adjuvant components mentioned above, comprise one or more pharmaceutically acceptable carriers or excipients, which include any excipient that does not itself induce the production of antibodies harmful to the individual receiving the composition.
  • Suitable excipients are typically large, slowly metabolised macromolecules such as proteins, saccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, sucrose, lactose and lipid aggregates (such as oil droplets or liposomes).
  • Such carriers are well known to those of ordinary skill in the art.
  • the vaccines may also contain diluents, such as water, saline, glycerol, etc.
  • auxiliary substances such as wetting or emulsifying agents, pH buffering substances, and the like, may be present.
  • Sterile pyrogen- free, phosphate buffered physiologic saline is a typical carrier.
  • a thorough discussion of pharmaceutically acceptable excipients is available in reference Gennaro, 2000, Remington: The Science and Practice of Pharmacy, 20th edition, ISBN:0683306472.
  • compositions of the present disclosure may be lyophilized or in aqueous form, i.e., solutions or suspensions. Liquid formulations of this type allow the compositions to be administered direct from their packaged form, without the need for reconstitution in an aqueous medium, and are thus ideal for injection.
  • Compositions may be presented in vials, or they may be presented in ready filled syringes. The syringes may be supplied with or without needles. A syringe will include a single dose of the composition, whereas a vial may include a single dose or multiple doses (e.g., 2 doses).
  • compositions of the present disclosure are also suitable for reconstituting other compositions from a lyophilized form.
  • the invention provides a kit, which may comprise two vials, or may comprise one ready-filled syringe and one vial, with the contents of the syringe being used to reconstitute the contents of the vial prior to injection.
  • Compositions of the present disclosure may be packaged in unit dose form or in multiple dose form (e.g., 2 doses). For multiple dose forms, vials are preferred to pre-filled syringes. Effective dosage volumes can be routinely established, but a typical human dose of the composition for injection has a volume of 0.5 mL.
  • compositions of the present disclosure may have a pH of between about 6.0 and about 8.0, in another embodiment, compositions of the invention have a pH of between 6.3 and 6.9, e.g., 6.6 ⁇ 0.2.
  • Compositions may be buffered at this pH. Stable pH may be maintained by the use of a buffer. If a composition comprises an aluminum hydroxide salt, a histidine buffer may be used. The composition should be sterile and/or pyrogen free.
  • compositions of the present disclosure may be isotonic with respect to humans.
  • compositions of the present disclosure may include an antimicrobial, particularly when packaged in a multiple dose format.
  • Antimicrobials may be used, such as 2- phenoxyethanol or parabens (methyl, ethyl, propyl parabens).
  • Any preservative is preferably present at low levels.
  • Preservative may be added exogenously and/or may be a component of the bulk antigens which are mixed to form the composition (e.g., present as a preservative in pertussis antigens).
  • compositions of the present disclosure may comprise a detergent, e.g., a TWEEN (polysorbate), such as TWEEN 80.
  • a detergent e.g., a TWEEN (polysorbate), such as TWEEN 80.
  • Detergents are generally present at low levels e.g. ⁇ 0.01%.
  • compositions of the present disclosure may include sodium salts (e.g., sodium chloride) to give tonicity.
  • the composition may comprise sodium chloride.
  • the concentration of sodium chloride in the composition of the invention is in the range of 0.1 to 100 mg/mL (e.g., 1-50 mg/mL, 2-20 mg/mL, 5-15 mg/mL) and in a further embodiment the concentration of sodium chloride may be 10 ⁇ 2 mg/mL NaCl e.g. about 9 mg/mL.
  • compositions of the present disclosure will generally include a buffer.
  • a phosphate or histidine buffer is typical.
  • compositions of the present disclosure may include free phosphate ions in solution (e.g., by the use of a phosphate buffer) in order to favor non-adsorption of antigens.
  • concentration of free phosphate ions in the composition of the invention is in one embodiment between 0.1 and 10.0 mM, or in another embodiment between 1 and 5 mM, or in a further embodiment about 2.5 mM.
  • compositions disclosed herein may be formulated into various dosage forms and administered by a number of different means that will deliver a therapeutically effective amount of the antigen or antibody.
  • Such compositions can be administered orally, parenterally, or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired.
  • Topical administration may also involve the use of transdermal administration such as transdermal patches or iontophoresis devices.
  • parenteral as used herein includes subcutaneous, intravenous, intramuscular, or intrasternal injection, or infusion techniques. Formulation of drugs is discussed in, for example, Gennaro, A. R., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.
  • a composition may be an intramuscular formulation.
  • Solid dosage forms for oral administration include capsules, tablets, caplets, pills, powders, pellets, and granules.
  • the active ingredient is ordinarily combined with one or more pharmaceutically acceptable excipients.
  • Oral preparations may also be administered as aqueous suspensions, elixirs, or syrups.
  • the active ingredient may be combined with various sweetening or flavoring agents, coloring agents, and, if so desired, emulsifying and/or suspending agents, as well as diluents such as water, ethanol, glycerin, and combinations thereof.
  • the preparation may be an aqueous or an oil- based solution.
  • Aqueous solutions may include a sterile diluent such as water, saline solution, a pharmaceutically acceptable polyol such as glycerol, propylene glycol, or other synthetic solvents; an antibacterial and/or antifungal agent such as benzyl alcohol, methyl paraben, chlorobutanol, phenol, thimerosal, and the like; an antioxidant such as ascorbic acid or sodium bisulfite; a chelating agent such as etheylenediaminetetraacetic acid; a buffer such as acetate, citrate, or phosphate; and/or an agent for the adjustment of tonicity such as sodium chloride, dextrose, or a polyalcohol such as mannitol or sorbitol.
  • a sterile diluent such as water, saline solution, a pharmaceutically acceptable polyol such as glycerol, prop
  • the pH of the aqueous solution may be adjusted with acids or bases such as hydrochloric acid or sodium hydroxide.
  • Oil-based solutions or suspensions may further comprise sesame, peanut, olive oil, or mineral oil.
  • the compositions may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carried, for example water for injections, immediately prior to use.
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.
  • compositions adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils.
  • the pharmaceutical composition is applied as a topical ointment or cream.
  • the active ingredient may be employed with either a paraffinic or a water-mi scible ointment base.
  • the active ingredient may be formulated in a cream with an oil-in-water cream base or a water-in-oil base.
  • compositions adapted for topical administration to the eye include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent.
  • Pharmaceutical compositions adapted for topical administration in the mouth include lozenges, pastilles and mouth washes.
  • Transmucosal administration may be accomplished through the use of nasal sprays, aerosol sprays, tablets, or suppositories, and transdermal administration may be via ointments, salves, gels, patches, or creams as generally known in the art.
  • an antigen or antibody of the invention is encapsulated in a suitable vehicle to either aid in the delivery of the antigen or antibody to target cells, to increase the stability of the composition, or to minimize potential toxicity of the composition.
  • a suitable vehicle is suitable for delivering a composition of the present invention.
  • suitable structured fluid delivery systems may include nanoparticles, liposomes, microemulsions, dendrimers and other phospholipid-containing systems. Methods of incorporating compositions into delivery vehicles are known in the art.
  • a liposome delivery vehicle may be utilized.
  • Liposomes are suitable for delivery of antigen or antibody in view of their structural and chemical properties.
  • liposomes are spherical vesicles with a phospholipid bilayer membrane.
  • the lipid bilayer of a liposome may fuse with other bilayers (e.g., the cell membrane), thus delivering the contents of the liposome to cells.
  • antigen may be selectively delivered to a cell by encapsulation in a liposome that fuses with the targeted cell's membrane.
  • Liposomes may be comprised of a variety of different types of phospholipids having varying hydrocarbon chain lengths.
  • Phospholipids generally comprise two fatty acids linked through glycerol phosphate to one of a variety of polar groups. Suitable phospholipids include phosphatidic acid (PA), phosphatidyl serine (PS), phosphatidylinositol (PI), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylcholine (PC), phosphatidylethanolamine (PE), and linear polyethylenimine (I-PEI).
  • the liposome may be comprised of linear polyethylenimine (I-PEI).
  • the fatty acid chains comprising the phospholipids may range from about 6 to about 26 carbon atoms in length, and the lipid chains may be saturated or unsaturated.
  • Suitable fatty acid chains include (common name presented in parentheses) n-dodecanoate (laurate), n-tetradecanoate (myristate), n-hexadecanoate (palmitate), n-octadecanoate (stearate), n-eicosanoate (arachidate), n-docosanoate (behenate), n-tetracosanoate (lignocerate), cis-9-hexadecenoate (palmitoleate), cis-9-octadecanoate (oleate), cis,cis-9, 12-octadecandienoate (linoleate), all cis- 9,12, 15-octadecatrienoate (linolenate
  • the two fatty acid chains of a phospholipid may be identical or different.
  • Acceptable phospholipids include dioleoyl PS, dioleoyl PC, distearoyl PS, distearoyl PC, dimyristoyl PS, dimyristoyl PC, dipalmitoyl PG, stearoyl, oleoyl PS, palmitoyl, linolenyl PS, and the like.
  • the phospholipids may come from any natural source, and, as such, may comprise a mixture of phospholipids.
  • egg yolk is rich in PC, PG, and PE
  • soy beans contains PC, PE, PI, and PA
  • animal brain or spinal cord is enriched in PS.
  • Phospholipids may come from synthetic sources too. Mixtures of phospholipids having a varied ratio of individual phospholipids may be used. Mixtures of different phospholipids may result in liposome compositions having advantageous activity or stability of activity properties.
  • phospholipids may be mixed, in optimal ratios with cationic lipids, such as N-(l-(2,3-dioleolyoxy)propyl)-N,N,N-trimethyl ammonium chloride, l,l '-dioctadecyl- 3,3,3 ',3'-tetramethylindocarbocyanine perchloarate, 3,3 '-deheptyloxacarbocyanine iodide, l,l '-dedodecyl-3,3,3',3'-tetramethylindocarbocyanine perchloarate, 1, 1 '-dioleyl-3,3,3 ',3'- tetramethylindo carbocyanine methanesulfonate, N-4-(delinoleylaminostyryl)-N- methylpyridinium iodide, or l, l,-dilinoleyl-3,
  • Liposomes may optionally comprise sphingolipids, in which sphingosine is the structural counterpart of glycerol and one of the one fatty acids of a phosphoglyceride, or cholesterol, a major component of animal cell membranes.
  • Liposomes may optionally contain pegylated lipids, which are lipids covalently linked to polymers of polyethylene glycol (PEG). PEGs may range in size from about 500 to about 10,000 Daltons.
  • PEGs polyethylene glycol
  • Liposomes may further comprise a suitable solvent.
  • the solvent may be an organic solvent or an inorganic solvent.
  • Suitable solvents include, but are not limited to, dimethylsulfoxide (DMSO), methylpyrrolidone, N-methylpyrrolidone, acetronitrile, alcohols, dimethylformamide, tetrahydrofuran, or combinations thereof.
  • DMSO dimethylsulfoxide
  • methylpyrrolidone methylpyrrolidone
  • N-methylpyrrolidone N-methylpyrrolidone
  • acetronitrile acetronitrile
  • alcohols dimethylformamide
  • tetrahydrofuran or combinations thereof.
  • Liposomes carrying antigen or antibody may be prepared by any known method of preparing liposomes for drug delivery, such as, for example, detailed in U.S. Pat. Nos. 4,241,046, 4,394,448, 4,529,561, 4,755,388, 4,828,837, 4,925,661, 4,954,345, 4,957,735, 5,043, 164, 5,064,655, 5,077,211 and 5,264,618, the disclosures of which are hereby incorporated by reference in their entirety.
  • liposomes may be prepared by sonicating lipids in an aqueous solution, solvent injection, lipid hydration, reverse evaporation, or freeze drying by repeated freezing and thawing.
  • the liposomes are formed by sonication.
  • the liposomes may be multilamellar, which have many layers like an onion, or unilamellar.
  • the liposomes may be large or small. Continued high-shear sonication tends to form smaller unilamellar lipsomes.
  • liposome formation may be varied. These parameters include, but are not limited to, temperature, pH, concentration of methionine compound, concentration and composition of lipid, concentration of multivalent cations, rate of mixing, presence of and concentration of solvent.
  • the present disclosure is directed to a method a method of treating a disease or condition in a subject.
  • the method comprises administering a therapeutically-effective amount of a triblock peptide to the subject, wherein the triblock peptide is of the formula: A - B - C wherein A is a lipid moiety; and B and C are independently a peptide block or a zwitterion-like block, including any of the peptides discussed above.
  • the triblock peptide is preferably administered in a pharmaceutical composition of the present invention, including any of the pharmaceutical compositions discussed above.
  • the pharmaceutical composition may be a vaccine composition of the present invention, including any of the vaccine compositions discussed above.
  • a therapeutically-effective amount of a triblock peptide may be administered to a subject. Administration is performed using standard effective techniques.
  • the disease or condition may be a pathogenic induced disease or condition, cancer or an autoimmune disease or condition.
  • autoimmune diseases may include, without limit, rheumatoid arthritis, multiple sclerosis, type 1 diabetes, lupus, celiac disease, Crohn's disease, ulcerative colitis, glomerulonephritis, chronic Lyme disease, Addison's disease, psoriasis, and scleroderma.
  • vasoactive intestinal peptide has distinct anti-inflammatory effects including downregulating T F- ⁇ by activated antigen presenting cells (APCs), specifically macrophages (M0s) and dendritic cells (DCs).
  • APCs activated antigen presenting cells
  • M0s macrophages
  • DCs dendritic cells
  • Dosages of the triblock peptide can vary between wide limits, depending on the disease or condition to be treated, the age of the subject, and the condition of the subject to be treated.
  • Duration of treatment could range from a single dose administered on a onetime basis to a life-long course of therapeutic treatments.
  • the duration of treatment can and will vary depending on the subject and the disease or disorder to be treated.
  • the duration of treatment may be for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days.
  • the duration of treatment may be for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks.
  • the duration of treatment may be for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months.
  • the duration of treatment may be for 1 year, 2 years, 3 years, 4 years, 5 years, or greater than 5 years.
  • administration may be frequent for a period of time and then administration may be spaced out for a period of time.
  • duration of treatment may be 5 days, then no treatment for 9 days, then treatment for 5 days.
  • the frequency of dosing may be once, twice, three times or more daily or once, twice, three times or more per week or per month, or as needed as to effectively treat the symptoms or disease.
  • the frequency of dosing may be once, twice or three times daily.
  • a dose may be administered every 24 hours, every 12 hours, or every 8 hours.
  • the frequency of dosing may be once, twice or three times weekly.
  • a dose may be administered every 2 days, every 3 days, or every 4 days.
  • the frequency of dosing may be one, twice, three or four times monthly.
  • a dose may be administered every 1 week, every 2 weeks, every 3 weeks, or every 4 weeks.
  • subject refers to, but are not limited to, a human, a livestock animal, a companion animal, a lab animal, and a zoological animal.
  • the subject may be a livestock animal.
  • the subject may be a bovine animal, a porcine animal, or a poultry animal.
  • the subject may be a cow, a pig, or a chicken.
  • suitable livestock animals may include pigs, cows, horses, goats, sheep, llamas, and alpacas.
  • the subject may be a companion animal.
  • companion animals may include pets such as dogs, cats, rabbits, and birds.
  • the subject may be a zoological animal.
  • the subject may be a human.
  • the human subj ect may be of any age. In some embodiments, the human subj ect may be about 20, about 25, about 30, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95 years of age or older. In some embodiments, the human subject is 30 years of age or older. In other embodiments, the human subject is 40 years of age or older. In other embodiments, the human subject is 45 years of age or older. In yet other embodiments, the human subject is 50 years of age or older. In still other embodiments, the human subject is 55 years of age or older. In other embodiments, the human subject is 60 years of age or older. In yet other embodiments, the human subject is 65 years of age or older.
  • the human subject is 70 years of age or older. In other embodiments, the human subject is 75 years of age or older. In still other embodiments, the human subject is 80 years of age or older. In yet other embodiments, the human subject is 85 years of age or older. In still other embodiments, the human subject is 90 years of age or older.
  • vaccine means a composition that when administered to a subject, typically elicits a protective immune response, where a protective immune response is one that ameliorates one or more symptoms of the target disorder.
  • treat or “treating” are meant to mean preventing or delaying an initial or subsequent occurrence of a disease or condition; increasing the disease-free survival time between the disappearance of a disease or condition and its reoccurrence; stabilizing or reducing an adverse symptom associated with a disease or condition; or inhibiting or stabilizing the progression of a disease or condition.
  • This includes prophylactic treatment, in which treatment before the disease or condition is established, prevents or reduces the severity or duration of the disease or condition.
  • the length of time a patient survives after being diagnosed with a disease or condition and treated using a method of the invention is at least 20, 40, 60, 80, 100, 200, or even 500% greater than (i) the average amount of time an untreated patient survives, or (ii) the average amount of time a patient treated with another therapy survives.
  • DIPEA N,N- diisopropylethylamine
  • Fmoc 9-fluorenylmethyloxycarbonyl
  • HBTU 2-(lH-benzotriazole-l- yl)-l, l,3,3-tetranethyluronium hexafluorophosphate
  • HOBt 1-hydroxybenzotriazole
  • HPLC high pressure liquid chromatography
  • LC-ESI-MS liquid chromatography-Electro Spray Ionization-mass spectrometry
  • TFA trifluoroacetic acid
  • Trt Trityl
  • TIS Triisopropylsilane
  • tBu t-butyl
  • Pbf 2,2,4,6,7-pentamethyl-dihydroben-zofurane-5-sulfonyl
  • Boc t-Butoxy.
  • EXAMPLE 1 Instructive Design of Triblock Peptide Amphiphil
  • Peptides were synthesized on Sieber amide resin (Chem-Impex International, SC Wood Dale, IL) by solid phase synthesis on a multiple peptide synthesizer (Advanced ChemTech 396 Omega, Louisville, KY) using Fmoc chemistry.
  • the peptide chain was assembled by sequential acylation (20 minute coupling) with in situ activated Fmoc amino acids. Recoupling was automatically performed at every cycle.
  • Fmoc amino acid activation was carried out using uronium salts (HBTU, 2.7 eq., HOBT 3 equiv) and DIEA (6 equiv).
  • Amino acid side chain protecting groups were tBu (Glu, Ser), Boc (Lys), Trt (Gin, His, Asn), and Pbf (Arg).
  • Fmoc protecting groups were removed at each amino acid addition cycle by treatment with 25% piperidine in dimethylformamide (DMF) for 15 minutes. Palmitic acid (Palm) tail modification was achieved by orthogonal deprotection with the aid of either Fmoc- Lys(Fmoc)-OH or Fmoc-Lys(Dde)-OH conjugated to the N terminus of the peptides depending on whether single or double fatty acid lipid conjugation was desired. Dde was removed by treating the peptide on resin with 2% Hydrazine in DMF.
  • Fmoc-Lys(Fmoc)-OH, Fmoc- Lys(Dde)-OH, and Palm conjugation were conducted manually in a glass reaction vessel (Chemglass, Vineland, NJ). All peptides were cleavage from resin and their side groups deprotected via a single-step reaction consisting of 2 hour exposure to the following mixture: TFA, thioanisole, phenol, water, ethandithiol and triisopropylsilane (87.5:2.5:2.5:2.5:2.5). Precipitation and multiple washing with diethyl ether yielded crude peptide product.
  • CMC Critical micelle concentration
  • TEM grids 200 mesh with standard thickness carbon support films were purchased from Electron Microscopy Sciences and glow discharged for 45 seconds (Pelco Easiglow) to impart a negative charge.
  • Product solutions (5 ⁇ .) were added to freshly glow- discharged films and incubated for 5 minutes. Filter paper was used to wick away excess solution and 5 ⁇ _, of nanotungsten (Nanoprobes, Inc.) was immediately added. After 5 minute of incubation, grids were blotted dried and stored for later use. Samples were imaged with a JEOL JEM- 1400 TEM at 120 kV for shape assessment.
  • Tilt series images were collected at 200 kV, spot size 4, gun lens of 5, and extraction voltage of 3950 sA at a nominal 23,000 x magnification with an underfocus of 1 sm.
  • Tilt increments were collected every 2 degrees with a tilt range of ⁇ 70°, starting at 0°, with the negative half of the tilt series collected using FEI Xplore3D.
  • Frames were aligned using FMOD with the patch-tracking algorithm using the entire imaged area for frame alignment and reconstructed with the weighted back projection algorithm.
  • Micellar diameters were measured using ImageJ software (NIH). Three different spots from each micelle were measured and the results from three separate micelles were averaged for each group.
  • Micelle secondary structure was investigated by circular dichroism using a circular dichroism spectrometer model 62DS (Aviv Biomedical, Inc., Lakewood, NJ). Micelle solutions (40 ⁇ ) were loaded into a 1 mm cuvette and measured a total of 10 times from 190 to 250 nm with an interval of 1 nm. The averaged data was curve fit using a linear combination of polylysine basis structures to calculate approximate a-helix, /3-sheet, and random coil content.
  • Peptide secondary structure was also affected by pH as evidenced by significant shifts in the sample CD spectra (FIG. 5D and FIG. 7 A) and corresponding secondary structure content estimation (FIG. 5E and FIG. 7B).
  • the zwitterion-like (KE) 4 block should possess a net positive charge at highly acidic pH, a near net zero charge at neutral pH, and a net negative charge at highly basic pH (FIG. 8).
  • Zeta potential measurement of the (KE) 4 peptide region alone at neutral pH was found to be slightly negative that became highly positively or negatively charged under acidic or basic conditions, respectively (Table 5), confirming the pH sensitivity of the region.
  • the alternating charge groups of (KE) 4 at neutral pH can create local dipole moments that allow for intermicellar complexation yielding the observed twining behavior.
  • twine-like micelles share some similar overall shape characteristics as previously reported ribbon-like micelles, those structures are individual micelles whose formation is governed by unevenly distributed hydrogen bonding and were found to be insensitive to changes in solution pH.
  • the hydrophobic region i.e. PalmK
  • OVABT-(KE) 4 the hydrophobic region was excluded (i.e., OVABT-(KE) 4 )
  • no ultrastructure structure was detected using TEM (data not shown) revealing that the hydrophobic driving force is vital to the micelle formation necessary to achieve twine-like ultrastructure.
  • PalmK-(EK) 4 -OVA BT individual micelles twined together like PalmK-OVABT-(KE) 4 , but self-assembled further by wrapping together three twines to form higher order structures at neutral pH up to tens of microns in length (FIG. 10B, FIG. 11 A, FIG. 11B, and FIG. 11C).
  • Peptide secondary structure was similarly affected by pH as evidenced by changes in the CD spectra (FIG. 10D) and secondary structure content (FIG. 10E). Therefore, similarly to twine-like micelles, electrostatic attractive forces between individual cylinders are believed to be the driving force for forming braid-like micelles at moderate to neutral pH.
  • PalmK-(EK) 4 -OVABT The diameter of the thread-like micelles in PalmK-(EK) 4 -OVABT were found to be similar in size to PalmK-OVA B T-(EK) 4 thread-like micelles (FIG. 12A and FIG. 12B and Table 6). This indicates that individual cylindrical micelles are the building blocks of both complex micellar structures. However, these formulations yielded quite different final ultrastructures at neutral pH. While both PAs are believed to undergo intermicellar dipole moment matching, PalmK-(EK)4-OVABT is thought to be unable to pair all of its surface dipole moments during the twining process (FIG. 13). These unmatched dipole moments participate in further electrostatic interactions by twisting together several twine-like micelles to form braid-like micelles.
  • PalmK-OVABT-(KE)4 PAs transition from very hydrophobic (PalmK) to modestly hydrophilic (OVABT) to highly hydrophilic ((KE) 4 ) likely aligning in this orientation from the core to the corona of the micelle allowing for significant PA mobility in the nanostructure.
  • PalmK-(EK)4-OVABT causes the PAs to bend the OVABT block back toward the core exposing the more hydrophilic (EK) 4 block on the corona yielding flower-like micelles.
  • PalmK-(EK)4-OVABT PAs less mobile and unable to rearrange in ways that fully charge match during the twining process therefore requiring further complexation through braiding to associate additional dipole moments.
  • This extra complexation means fewer charge matches between each twine yielding overall weaker intermolecular forces holding them together.
  • This theory is corroborated by the less extreme pH changes (i.e., 3/ 11 versus 2/12) required to break apart the individual micelles and the more significant extent of dissociation seen for PalmK-(EK) 4 -OVA B T PAMs (FIG. 10A, FIG. 10B, FIG. IOC, FIG. 10D, and FIG. 10E) compared to PalmK-OVA B T-(KE) 4 PAMs (FIG. 5A, FIG. 5B, FIG. 5C, FIG. 5D, and FIG. 5E)
  • PalmK-(EK)4-OVABT thread-like micelles were occasionally observed as small fibers at neutral pH (FIG. 11 A, FIG. 11B, and FIG. 11C). This indicates the existence of different types of ⁇ -sheet formation due to different orientations of carbonyl- amide stretching. Thread-like micelle fibers form when hydrogen bonding occurs in two parallel planes, in which case the stretching angle is 0°.
  • PalmK-OVABT- (KE)4 and PalmK-(EK)4-OVABT were dissolved in micelles in either neutral pH corrected milli-Q double distilled water (ddH 2 0) or phosphate buffered saline (PBS). No significant differences were observed for either morphology (FIG. 14A, FIG. 14B, FIG. 14C, and FIG.
  • the PA hydrophobic moiety has been found to play an important role in micellization, impacting the resulting nanomaterial properties.
  • two different hydrophobic blocks possessing either one (PalmK) or two (Palm 2 K) palmitic acid tails were tested.
  • the ability to change the number of hydrophobic tails was achieved by orthogonally protecting the N-terminal non-native lysine so that either one or two palmitic acid tails could be conjugated.
  • Adding a second tail was found to dramatically alter micellar morphology, both for micelles that with a zwitterion-like region (FIG. 16A, FIG. 16B, FIG. 16C, and FIG.
  • FIG. 16C created by its single palmitic acid counterpart (i.e., PalmK-(EK)4-OVABT) to cluster micelles (FIG. 16D).
  • PalmK-(EK)4-OVABT single palmitic acid counterpart
  • FIG. 16D cluster micelles
  • the two hydrophobic tail products showed minimal morphological (FIG. 18A, FIG. 18B, FIG. 18C, FIG. 18D, FIG. 18E, and FIG. 18F) and secondary structure (FIG. 19A, FIG. 19B, FIG. 19C, and FIG. 19D) changes due to pH-controlled zwitterion-like block charge modification. Slight aggregation was observed for Palm 2 K-OVABT-(KE)4 and significant aggregation was seen for Palm 2 K-(EK) 4 -OVA B T at neutral pH (FIG. 20A and FIG.
  • Palm 2 K-OVABT like PalmK-OVABT, showed no sensitivity to pH adjustment as no electrostatically sensitive region was included in this formulation (FIG. 21A, FIG. 21B, and FIG. 21C).
  • Vasoactive intestinal peptide is a 28-amino acid neuropeptide that has distinct anti-inflammatory effects including downregulating T F- ⁇ production by activated antigen presenting cells (APCs), specifically macrophages (MOs) and dendritic cells (DCs). It has also been shown to induce DCs to secrete CCL22 which recruit regulatory T cells (T reg s) that can facilitate localized tolerance.
  • APCs activated antigen presenting cells
  • MOs macrophages
  • DCs dendritic cells
  • T reg s regulatory T cells
  • VIP amphiphiles were created to investigate their capacity to form micelles (VIPAMs) and potentiate the bioactivity of VIP.
  • VIPAMs micelles
  • VIPA chemistries were produced.
  • the first VIPA was synthesized by directly conjugating palmitic acid (Palm) to the N-terminus of VIP to form Palm-VIP, HSDAVFTDNYTRLRKQMAVKKYLNSILN (SEQ ID NO: 3), (pVIPA - FIG. 39A).
  • the second VIPA included a zwitteri on-like peptide region between Palm and VIP yielding PalmK- (EK) 4 -VIP, KEKEKEKEKHSDAVFTDNYTRLRKQMAVKKYLNSILN (SEQ ID NO: 4) (pzVIPA - FIG. 39B).
  • Micellization of each VIPA was characterized using a critical micelle (CMC) assay and negative-stain aided transmission electron microscopy (TEM).
  • CMC critical micelle
  • TEM negative-stain aided transmission electron microscopy
  • Tumor necrosis factor alpha is a monocyte-derived cytokine that plays a significant role in the inflammatory response.
  • TNF-a is produced by M0s and DCs that are activated during infection, commonly due to the cell- based identification of pathogen associated molecule patterns, most notably lipopolysaccharide (LPS) found in the cell wall of gram-negative bacteria.
  • LPS lipopolysaccharide
  • Excessive TNF-a production has been shown to cause tissue injury, fever, atherosclerosis, and even death.
  • activated DCs tend to migrate to nearby lymph nodes where they activate naive T helper cells.
  • Activated effector T cells will migrate back to the inflammation site where they will recruit natural killer cells and additional M0s which further exacerbate the inflammatory response.
  • the B7 ligand CD86 present on activated DCs plays an important role in this cascade acting as a co-stimulatory signal for T cell activation. A lack of co-stimulatory signaling often leads to T cell anergy. Conversely, the presence of CD86 on DCs without corresponding MHC II antigen presentation plays a role in T re g induction.
  • pVIPA was unable to enhance the TNF-a suppressive effects of VIP in activated M0s (FIG. 41A) and completely nullified VIP effects on activated DC TNF-a secretion (FIG. 41B).
  • the only statistically significant anti-inflammatory effect for pVIPA over VIP was found in DC CD86 expression at the high concentration where it was actually enhanced (FIG. 41C).
  • pzVIPA nearly completely abrogated TNF-a secretion in activated M0s (FIG. 41A), maintained VIP -based TNF-a secretion in activated DCs (FIG. 41B), and significantly limited CD86 surface expression on activated DCs (FIG. 41C).
  • these enhancement effects were only observed at the high concentration and not at the low concentration.
  • T re gS are a unique type of suppressor T cell that facilitates peripheral immunological tolerance. Increasing the presence and development of T reg s at the effector site of autoimmunity or inflammation has been suggested as a potential treatment for immune-related disorders or transplant rejection. T re g recruitment to the desirable tissue site can be guided by the presence of a gradient of the chemokine CCL22 (MDC).
  • MDC chemokine CCL22
  • CD86 ligand presented by DCs is an important molecule that has been shown to induce T reg survival and expansion in peripheral tissue, especially in the absence of corresponding MHC II-presented antigen. Therefore, the enhanced expression of CD86 is a potential key factor that affects downstream immunoregulatory functions of CCL22-recruited T reg s. It was discovered that high concentration pVIPA induced the highest CD86 expression of VIP treated groups or lipid control groups for both mature DCs (FIG. 32C) and immature DCs (FIG. 42A, FIG. 42B, FIG. 42C, FIG. 42D, FIG. 42E, FIG. 42F, and FIG. 43C). The CCL22 and CD86 data suggest that cylindrical VIPAMs possess potential immunoregulatory properties.
  • VIP AM Structure-Bioactivity Relationships Interestingly, VIP is known to modulate TNF-a, CD86, and CCL22 expression through the same receptor ⁇ i.e. VPAC1), indicating that formulation chemistry and structure is very directly impacting peptide bioactivity.
  • VIP/VP AC interactions are known to be dependent on a number of factors including VIP concentration, amino acid availability, and conformation.
  • VIP concentration One of the major differences with peptide amphiphiles compared to peptides is their capacity to enhance peptide-cell interactions due to their lipid content. Therefore, both pVIPA and pzVIPA are expected to yield greater VIP concentrations at the cell surface.
  • the N-terminal amino acid of VIP ⁇ i.e.
  • histidine is known to play an important role in VIP/VP AC binding affinity. With the N-terminal histidine on pVIPA being directly lipidated, it is likely to be closer to the membrane and more rigid than pzVIPA which possesses a somewhat flexible linker ⁇ i.e. (KE) 4 ) between the lipid and VIP. Previous studies have demonstrated VIP a -helicity enhances peptide association with VP AC. Interestingly, CD analysis revealed that pzVIPA had more abundant a -helical conformation than both VIP and pVIPA (FIG. 41A and FIG. 41B). These factors may impact interactions between pzVIPA and VP AC leading to the enhanced TNF-a and CD86 suppression activity observed (FIG. 42A, FIG. 42B, and FIG. 42C).
  • DCs (FIG. 43A), the magnitude of this response was significantly different.
  • DC CCL22 induction is related to different factors including VIP/VP AC engagement and DC activation state.
  • pzVIPA is likely engaging VP AC, but without activating the DCs as no increase in CD86 cell surface expression was detected (FIG. 43C).
  • high concentration pVIPA significantly increased CD86 expression in immature DCs (FIG. 43C) without altering other stimulatory markers like CD40 expression and TNF-a production ⁇ data not shown) providing evidence of a semi-mature DC state similar to previous work exploring VIP.
  • This VTP- stimulated DC phenotype was found to directly correspond to more elevated levels of CCL22 production.
  • An across the board increase in CCL22 production for mature DCs (FIG.
  • Influenza is a common and highly communicable respiratory disease whose impact varies from year to year but is always incredibly significant as evidenced by data generated by the United States Center for Disease Control.
  • the influenza virus was estimated to be responsible for 9.2 million seasonal illnesses, 4.3 million medical visits, and 139,000 hospitalizations, numbers that are on the low end for seasonal disease.
  • the following season i.e. October 2012 - April 2013
  • This infectious disease is not only responsible for significant losses in productivity, but can also be quite lethal as evidenced by the approximately 291,000 - 646,000 worldwide deaths influenza is responsible for each year.
  • the highly mutable nature of the influenza virus can also yield a pathogenic pandemic strain that can emerge at any time similar to the one that was responsible for more than 500 million illnesses and 50 - 100 million deaths worldwide from 1918 to 1920.
  • influenza vaccines were able to be mass produced and used to vaccinate the armed forces followed by immunizing the general public. 4
  • Considerable efforts to improve the influenza vaccine have been made, the most significant of which focused on the development of an intranasally-delivered, live attenuated vaccine. In specific, presenting a weakened version of the virus directly to the primary site of infection (i.e. respiratory track) is hoped to be able to induce more durable and completely protective host immunity.
  • HlNl subtype influenza While fortunate, genetic recombination of HlNl subtype influenza within an animal vector (e.g. avian or swine) could lead to a mutated influenza strain with a high transmission rate and significant lethality in the near future.
  • an animal vector e.g. avian or swine
  • the criteria of such a vaccine would include the capacity to be highly effective against symptomatic infections, protect against both groups of influenza A, and last at least a year.
  • PAs peptide amphiphiles
  • peptide amphiphile micelles have been shown capable of increasing localized concentration, enhancing intracellular delivery, controlling peptide secondary structure, and facilitating cell-specific targeting which have been shown to synergistically lead to self-adjuvanting immune responses to B cell and CD8 T cell epitopes.
  • peptide antigens are often delivered with adjuvants.
  • aluminum salts were the only adjuvants FDA cleared for human use until monophosphoryl lipid A (MPLA) and Squalene/a-Tocopherol/polysorbate80 (AS03) were cleared as adjuvants for the human papillomavirus vaccine in 2009 and the H5N1 influenza virus vaccine in 2012, respectively.
  • AS03 is an oil-in-water emulsion adjuvant that has been profoundly effective in inducing highly immunogenic, long-lasting immune responses to influenza, but is believed to be responsible for increased incidents of narcolepsy among children receiving the adjuvant.
  • MPLA is the non-toxic portion of the bacterial endotoxin lipopolysaccharide (LPS) which has been found to be a Toll-like receptor-4 (TLR-4) agonist.
  • LPS bacterial endotoxin lipopolysaccharide
  • TLR-4 Toll-like receptor-4
  • MPLA is hydrophobic allowing for it to be readily entrapped within the peptide amphiphile micelle (PAM) core.
  • TLR agonists such dipalmitoylglycerylcysteinylserinyltetralysine (P2CSK4) for TLR-2, polyriboinosinic:polyriboctidylic acid (poly(LC)) for TLR-3, and CpG oligodeoxynucleotides (CpG ODNs) for TLR-9 are attractive molecular adjuvant candidates as well.
  • Hydrophobic P 2 C can be entrapped within the PAM core similarly to MPLA whereas negatively-charged poly(LC) and CpG ODN can be complexed to short positively-charged oligolysine repeats (i.e. K 8 ).
  • K 8 positively-charged oligolysine repeats
  • Grzybowski B. A. Electrostatic self-assembly of binary nanoparticle crystals with a diamond-like lattice. Science 2006, 312 (5772), 420-424.
  • amphiphiles with different hydrophobic blocks for simultaneous delivery of drugs and genes Macromol. Rapid Commun. 2010, 31 (13), 1212-1217.
  • Nonionic triblock and star diblock copolymer and oligomeric surfactant syntheses of highly ordered, hydrothermally stable, mesoporous silica structures J. Am. Chem. Soc. 1998, 120 (24), 6024-6036.
  • alternating lysines can act as a highly specific Z-DNA binding domain.
  • nanoparticles conjugated with a targeting peptide ligand selected by phage display technique are conjugated with a targeting peptide ligand selected by phage display technique. Macromolecular Bioscience 2015, 15 (3), 395-404.
  • cationic pepitde LL-37 binds Mac-1 (CD1 lb/CD18) with a low dissociation rate and promotes phagocytosis.
  • intestinal peptide receptor type 2 receptor deficient mice develop exacerbated experimental autoimmuno encephalomyelitis with increased Thl/Thl7 and reduced Th2/Treg responses. Brain, Behavior, and Immunity 2015, 44, 167-175.
  • Thermosensitive Nanoparticles with Degradable Cross-Links Suppresses Proinflammatory Cytokine Production. Biomacromolecules 2015, 16 (4), 1191-1200.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Zoology (AREA)
  • Toxicology (AREA)
  • Virology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Vascular Medicine (AREA)
  • Endocrinology (AREA)
  • Oncology (AREA)
  • Pulmonology (AREA)
  • Communicable Diseases (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

Selon un aspect, la présente invention concerne des peptides triblocs comprenant un fragment lipidique, un bloc peptidique et un bloc zwitterionique. Selon un autre aspect, l'invention concerne des compositions pharmaceutiques comprenant les peptides triblocs selon la présente invention agencés dans des micelles dans un véhicule pharmaceutiquement acceptable. Dans certains modes de réalisation, les compositions pharmaceutiques selon la présente invention sont des compositions vaccinales, qui peuvent en outre comprendre un adjuvant. Des méthodes d'utilisation des peptides triblocs et des compositions selon l'invention pour traiter une maladie ou une affection sont en outre décrites.
PCT/US2018/058200 2017-10-30 2018-10-30 Amphiphiles peptidiques triblocs, micelles et leurs méthodes d'utilisation WO2019089584A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US16/760,329 US20200339661A1 (en) 2017-10-30 2018-10-30 Triblock peptide amphiphiles, micelles and methods of use

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201762578843P 2017-10-30 2017-10-30
US62/578,843 2017-10-30

Publications (2)

Publication Number Publication Date
WO2019089584A2 true WO2019089584A2 (fr) 2019-05-09
WO2019089584A3 WO2019089584A3 (fr) 2019-06-13

Family

ID=66332906

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2018/058200 WO2019089584A2 (fr) 2017-10-30 2018-10-30 Amphiphiles peptidiques triblocs, micelles et leurs méthodes d'utilisation

Country Status (2)

Country Link
US (1) US20200339661A1 (fr)
WO (1) WO2019089584A2 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023049791A1 (fr) * 2021-09-22 2023-03-30 The Trustees Of Columbia University In The City Of New York Systèmes d'administration de lipides bioactifs

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7371719B2 (en) * 2002-02-15 2008-05-13 Northwestern University Self-assembly of peptide-amphiphile nanofibers under physiological conditions
AU2003298647A1 (en) * 2002-11-14 2004-06-15 Claussen, Randal, C. Synthesis and self-assembly of abc triblock bola peptide
US11286489B2 (en) * 2016-02-11 2022-03-29 The Curators Of The University Of Missouri POSH inhibitor complex biomolecules and amphiphile micelles

Also Published As

Publication number Publication date
WO2019089584A3 (fr) 2019-06-13
US20200339661A1 (en) 2020-10-29

Similar Documents

Publication Publication Date Title
US20120064110A1 (en) Synthetic nanocarrier vaccines comprising peptides obtained or derived from human influenza a virus hemagglutinin
JP6324068B2 (ja) 合成ナノキャリア混合ワクチン
Barrett et al. Modular peptide amphiphile micelles improving an antibody-mediated immune response to Group A Streptococcus
Siebert et al. Tuftsin-properties and analogs
US10245319B2 (en) Lymph node-targeting nanoparticles
Nevagi et al. Structure-activity relationship of group A streptococcus lipopeptide vaccine candidates in trimethyl chitosan-based self-adjuvanting delivery system
EP2598519A1 (fr) Agent antiviral
JP6158097B2 (ja) 炎症を抑制するためのペプチド
US20210106652A1 (en) Immunogenic Compounds For Treatment Of Fibrosis, Autoimmune Diseases And Inflammation
US20230090311A1 (en) Self-assembling, self-adjuvanting system for delivery of vaccines
Debnath et al. Potential application of bionanoparticles to treat severe acute respiratory syndrome coronavirus-2 infection
US20200339661A1 (en) Triblock peptide amphiphiles, micelles and methods of use
AU2018284219A1 (en) Hydrochloride salts of C5a receptor agonist peptides
JPH08506592A (ja) ３−ｏ−脱アシル化モノホスホリル脂質Ａ含有のインフルエンザワクチン組成物
WO2023141679A1 (fr) Dimères et leurs utilisations
WO2018038168A1 (fr) Peptide de liaison à l'hémagglutinine et agent prophylactique/thérapeutique pour une infection par le virus de la grippe qui contient celui-ci
JP2018070571A (ja) 免疫誘導剤
CN113813375A (zh) 一种新型抗新冠病毒复合物的组成及其在防治冠状病毒感染疾病药物中的应用
WO2010129710A1 (fr) Utilisation d'un rankl pour induire une différenciation des cellules microfold (cellules m)
US20230365647A1 (en) Mannose receptor-derived peptides for neutralizing pore-forming toxins for therapeutic uses
Santhosh et al. Nanovaccinology and superbugs
WO2022174768A1 (fr) Peptides anti-viraux et compositions et procédés d'utilisation associés
CN117015402A (zh) 抗病毒肽及其组合物和使用方法
Ruseska et al. Protamine–a review on an oligonucleotide-binding peptide applied in nanopharmaceuticals including vaccines
WO2023049921A1 (fr) Peptide inhibiteur de pd-l1 pour l'immunothérapie anticancéreuse

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18873578

Country of ref document: EP

Kind code of ref document: A2